Molecular basis of inheritane ppt
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Jul 06, 2014
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About This Presentation
Grade-12 Biology (CBSE)
Slide Content
Introduction
•Name the two type of nucleic acid found in living organism.
Deoxyribonucleic acid (DNA) & Ribonucleic acid (RNA)
•What act as the genetic material in most of the living organism?
DNA
•In virus what act as the genetic material other than DNA?
RNA
•List some functions of RNA
Messenger, catalytic molecule
•Name the two type of nucleic acid found in living organism.
Deoxyribonucleic acid (DNA) & Ribonucleic acid (RNA)
•What act as the genetic material in most of the living organism?
DNA
•In virus what act as the genetic material other than DNA?
RNA
•List some functions of RNA
Messenger, catalytic molecule
•Nucleic acids.
•Nucleic acids are the macromolecules present in all living cell.
•FreidrichMiescherwas the first person isolated the nucleic acids
from the pus cells. He called it as nuclein.
•As it has an acidic nature, hence Altmanncalled it as nucleic acids.
•The two types of nucleic acids found in living organisms are,
1.Deoxyribonucleic acid [DNA]
2.Ribonucleic acid [RNA].
•Nucleic acids are the macromolecules present in all living cell.
•FreidrichMiescherwas the first person isolated the nucleic acids
from the pus cells. He called it as nuclein.
•As it has an acidic nature, hence Altmanncalled it as nucleic acids.
•The two types of nucleic acids found in living organisms are,
1.Deoxyribonucleic acid [DNA]
2.Ribonucleic acid [RNA].
Deoxyribonucleic acid [DNA]
•Genetic material of all living organisms-carries the coded information
from one generation to another generation.
•In eukaryotic cell DNA is found nucleus. It is the chief component of
chromosomes.
•DNA also found in the mitochondria and chloroplasts
•Long polymer of deoxyribonucleotides.
•Length of the DNA -number of nucleotides or pair of nucleotide (base
pairs)
•Eg. Bacteriophage-ɸ 174-5386 nucleotides, Bacteriophage lambda-
48502 base pair (bp), human DNA-3.3 X 10
9
bp.
•Genetic material of all living organisms-carries the coded information
from one generation to another generation.
•In eukaryotic cell DNA is found nucleus. It is the chief component of
chromosomes.
•DNA also found in the mitochondria and chloroplasts
•Long polymer of deoxyribonucleotides.
•Length of the DNA -number of nucleotides or pair of nucleotide (base
pairs)
•Eg. Bacteriophage-ɸ 174-5386 nucleotides, Bacteriophage lambda-
48502 base pair (bp), human DNA-3.3 X 10
9
bp.
•DNA & RNA -polynucleotide chain
•A nucleotide –3 components
1.A nitrogenous base
2.A pentose sugar (DNA-deoxyribose sugar, RNA-Ribose sugar)
3.A phosphate group
•Nitrogenous base-Nitrogen containing compound
•Two types-Purines (Adenine & Guanine)&Pyrimidines (Cytosine, Uracil
& Thymine)
•Adenine, Guanine & Cytosine-Both DNA & RNA
•Thymine-only in DNA
•Uracil-Only in RNA in place of Thymine
•Nitrogenous Base linked is linked to pentose sugar through N-glycosidic
linkage-Nucleoside
Structure of polynucleotide chain
•A nucleotide –3 components
1.A nitrogenous base
2.A pentose sugar (DNA-deoxyribose sugar, RNA-Ribose sugar)
3.A phosphate group
•Nitrogenous base-Nitrogen containing compound
•Two types-Purines (Adenine & Guanine)&Pyrimidines (Cytosine, Uracil
& Thymine)
•Adenine, Guanine & Cytosine-Both DNA & RNA
•Thymine-only in DNA
•Uracil-Only in RNA in place of Thymine
•Nitrogenous Base linked is linked to pentose sugar through N-glycosidic
linkage-Nucleoside
Structure of polynucleotide chain
•Nucleoside-: compounds formed by the combination of Pentose sugar
and Nitrogenous base
•In DNA molecule-Deoxyribose sugar combines with nitrogenous base &
form Deoxyribonucleoside
1.Adenine + deoxyribose sugar = Deoxyadenosine
2.Guanine + deoxyribose sugar = Deoxygaunosine.
3.Cytosine + deoxyribose sugar = Deoxycytidine.
4.Thymine + deoxyribose sugar =Deoxythymidine
and Nitrogenous base
•In DNA molecule-Deoxyribose sugar combines with nitrogenous base &
form Deoxyribonucleoside
1.Adenine + deoxyribose sugar = Deoxyadenosine
2.Guanine + deoxyribose sugar = Deoxygaunosine.
3.Cytosine + deoxyribose sugar = Deoxycytidine.
4.Thymine + deoxyribose sugar =Deoxythymidine
•Nucleotides:The compound formed by the combination of phosphate
groupswith nucleosides is called nucleotides.
•Phosphate group is linked to nucleoside through 5’-OH of nucleoside
through phosphoesterlinkage-corresponding nucleotides
(deoxyridonucleotide)
•The four types of nucleotides present in the DNA are,
1.Deoxyadenosinemonophosphate.
2.Deoxygaunosinemonophosphate.
3.Deoxycytidinemonophosphate.
4.Deoxythymidinemonophosphate.
groupswith nucleosides is called nucleotides.
•Phosphate group is linked to nucleoside through 5’-OH of nucleoside
through phosphoesterlinkage-corresponding nucleotides
(deoxyridonucleotide)
•The four types of nucleotides present in the DNA are,
1.Deoxyadenosinemonophosphate.
2.Deoxygaunosinemonophosphate.
3.Deoxycytidinemonophosphate.
4.Deoxythymidinemonophosphate.
Polynucleotide strand:
•Two nucleotides are linked through 3’-5’ phosphodiesterlinkage-
dinucleotide
•The phospho-di-ester bond forms between 5th and 3rd carbon atom
position of pentose sugar. Therefore polynucleotide strand contains 5th
and 3rd end.
•More nucleotides joins-polynucleotide, which has at one end free
phosphate moiety at 5’ end -5’-end of polynucleotide & free 3’-OH
group-referred as 3’-end of polynucleotide
•Sugar & phosphate-backbone & Nitrogenous base project form the
backbone
•Two nucleotides are linked through 3’-5’ phosphodiesterlinkage-
dinucleotide
•The phospho-di-ester bond forms between 5th and 3rd carbon atom
position of pentose sugar. Therefore polynucleotide strand contains 5th
and 3rd end.
•More nucleotides joins-polynucleotide, which has at one end free
phosphate moiety at 5’ end -5’-end of polynucleotide & free 3’-OH
group-referred as 3’-end of polynucleotide
•Sugar & phosphate-backbone & Nitrogenous base project form the
backbone
Structure of rna
•Structure of RNA is similar to DNA
•Nucleotide residue has additional –OH group
•-OH group is present at 2’-position in the ribose
•Nitrogenous base Uracil is found in place of Thymine
•Reads the template of DNA and forms the protein required for the
cell
•Structure of RNA is similar to DNA
•Nucleotide residue has additional –OH group
•-OH group is present at 2’-position in the ribose
•Nitrogenous base Uracil is found in place of Thymine
•Reads the template of DNA and forms the protein required for the
cell
History-Structure of DNA-polynucleotide
•James Watson and Francis Crick first
proposed the structural model of DNA in 1953
based on the X-ray diffraction data-Maurice
Wilkins & Rosalind Franklin
•They got the Nobel Prize for their work in
1962.
•Proposed Double Helix model for structure of
DNA-remarkable proposition was base
pairing between two strand of polynucleotide
•Proposition depend on observation of Erwin Chargaff that for a double
stranded DNA, the ratios between Adenine & Thymine ; Guanine &
Cytosine are constant & equal
•Base pairing-unique property of polynucleotide chain, complementary to
each other-sequence of one DNA strand known then sequence of other
strand can be predicted
•Each strand of DNA (parental DNA)-template for synthesis of new strand-
two double stranded DNA (daughter DNA) will be identical to parental
DNA molecule
•James Watson and Francis Crick first
proposed the structural model of DNA in 1953
based on the X-ray diffraction data-Maurice
Wilkins & Rosalind Franklin
•They got the Nobel Prize for their work in
1962.
•Proposed Double Helix model for structure of
DNA-remarkable proposition was base
pairing between two strand of polynucleotide
•Proposition depend on observation of Erwin Chargaff that for a double
stranded DNA, the ratios between Adenine & Thymine ; Guanine &
Cytosine are constant & equal
•Base pairing-unique property of polynucleotide chain, complementary to
each other-sequence of one DNA strand known then sequence of other
strand can be predicted
•Each strand of DNA (parental DNA)-template for synthesis of new strand-
two double stranded DNA (daughter DNA) will be identical to parental
DNA molecule
The structure of DNA is as follows:
•Two polynucleotide chains-the backbone of polynucleotide is formed
by sugar-phosphate & bases project inside
•Two chains: anti-parallel polarity; one chain has polarity 5’3’ &
other chain 3’5’
•Bases of two strand projecting inside-paired through hydrogen bond
(H-bond) & form base pairs (bp)
•Adenine form two hydrogen bonds (H-bond) with Thymine & Guanine
form three H-bond with Cytosine
•Purines comes opposite with Pyrimidines-generates uniform distance
between two strands of the helix
Adenine Thymine
Guanine Cytosine
•Two polynucleotide chains-the backbone of polynucleotide is formed
by sugar-phosphate & bases project inside
•Two chains: anti-parallel polarity; one chain has polarity 5’3’ &
other chain 3’5’
•Bases of two strand projecting inside-paired through hydrogen bond
(H-bond) & form base pairs (bp)
•Adenine form two hydrogen bonds (H-bond) with Thymine & Guanine
form three H-bond with Cytosine
•Purines comes opposite with Pyrimidines-generates uniform distance
between two strands of the helix
Adenine Thymine
Guanine Cytosine
•Two chains-coiled right handed fashion
•Pitch of helix is 3.4 nm-Thepitchof a helix is the width of one complete
helix turn, measured parallel to the axis of the helix.
•10 bpin each turn or pitch, distance between a bpin helix-0.34nm
•Plane of one base pair stacks over the otherin double helix-addition to H-
bond confers stability
•Stability of DNA structure : H-bond & stacking of nitrogenous base
AdenineThymine
GuanineCytosine
•Pitch of helix is 3.4 nm-Thepitchof a helix is the width of one complete
helix turn, measured parallel to the axis of the helix.
•10 bpin each turn or pitch, distance between a bpin helix-0.34nm
•Plane of one base pair stacks over the otherin double helix-addition to H-
bond confers stability
•Stability of DNA structure : H-bond & stacking of nitrogenous base
AdenineThymine
GuanineCytosine
•Francis Crick proposed Central dogmain molecular biology
•Genetic information flows from DNARNAProtein
•The coded genetic information hard-wired into DNA is transcribed into
individual transportable cassettes, composed of messenger RNA
(mRNA); each mRNA cassette contains the program for synthesis of a
particular protein (or small number of proteins)
DNA
mRNA
Protein
Transcription
Translation
•Genetic information flows from DNARNAProtein
•The coded genetic information hard-wired into DNA is transcribed into
individual transportable cassettes, composed of messenger RNA
(mRNA); each mRNA cassette contains the program for synthesis of a
particular protein (or small number of proteins)
DNA
mRNA
Protein
Transcription
Translation
1. Molecules of DNA are composed of long chains of
•amino acids
•fatty acids
•monosaccharides
•nucleotides
Ans.-Nucleotides
2. The primary function of DNA is to
•make proteins
•store and transmit genetic information
•control chemical processes within cells
•prevent mutations
Ans.-Make protein
Lets Practice-dna structure
•amino acids
•fatty acids
•monosaccharides
•nucleotides
Ans.-Nucleotides
2. The primary function of DNA is to
•make proteins
•store and transmit genetic information
•control chemical processes within cells
•prevent mutations
Ans.-Make protein
Lets Practice-dna structure
3. A nucleotide consists of
•a sugar, a protein, and adenine
•a sugar, an amino acid, and a starch
•a sugar, a phosphate group, and a nitrogen-containing base
•a starch, a phosphate group, and a nitrogen-containing base
Ans.-A sugar, a phosphate group and a nitrogen containing base
4. The part of the molecule for which deoxyribonucleic acid is named
is
•phosphate group
•sugar
•nitrogen base
•none of the above
Ans.-Sugar
•a sugar, a protein, and adenine
•a sugar, an amino acid, and a starch
•a sugar, a phosphate group, and a nitrogen-containing base
•a starch, a phosphate group, and a nitrogen-containing base
Ans.-A sugar, a phosphate group and a nitrogen containing base
4. The part of the molecule for which deoxyribonucleic acid is named
is
•phosphate group
•sugar
•nitrogen base
•none of the above
Ans.-Sugar
5. Purines and pyrimidinesare
•bases found in amino acids
•able to replace phosphate groups from defective DNA
•names of specific types of DNA molecules
•bases found in nucleotides
Ans.-Bases found in nucleotides
6. The scientists credited with determining the structure of DNA are
•Avery and Chargaff
•Hershey and Chase
•Mendel and Griffith
•Watson and Crick
Ans.-Watson and Crick
•bases found in amino acids
•able to replace phosphate groups from defective DNA
•names of specific types of DNA molecules
•bases found in nucleotides
Ans.-Bases found in nucleotides
6. The scientists credited with determining the structure of DNA are
•Avery and Chargaff
•Hershey and Chase
•Mendel and Griffith
•Watson and Crick
Ans.-Watson and Crick
7. The base-pairing rules state that the following are base pairs in
DNA:
•adenine -thymine; guanine -cytosine
•adenine -thymine; uracil -cytosine
•adenine -guanine; thymine -cytosine
•uracil -thymine; guanine –cytosine
Ans.-Adenine-Thymine; Guanine-Cytosine
8. ATTG : TAAC ::
•AAAT : TTTG
•TCGG : AGAT
•GTCC : CAGG
•CGAA : TGCG
Ans.-GTCC: CAGG
DNA:
•adenine -thymine; guanine -cytosine
•adenine -thymine; uracil -cytosine
•adenine -guanine; thymine -cytosine
•uracil -thymine; guanine –cytosine
Ans.-Adenine-Thymine; Guanine-Cytosine
8. ATTG : TAAC ::
•AAAT : TTTG
•TCGG : AGAT
•GTCC : CAGG
•CGAA : TGCG
Ans.-GTCC: CAGG
Packaging of dna helix
•Length of typical Double DNA helix in mammalian calculated as 2.2
meters
•Cells-Eukaryotes & Prokaryotes
•Eukaryotes DNA localized inside the nucleus
•Prokaryotes like E. Coli-defined nucleus & DNA is not scattered
throughout in cell
•DNA-negatively charged; Protein-Positive charges
•DNA held with positive charged Protein-region termed ‘Nucleoid’
•DNA in nucleoid organized in large loops held by protein
•Eukaryotes-organization is complex; DNA forms complex with
positively charged protein-histones
•Proteins acquires charge-abundance of amino acid residues lysines&
arginines
•Amino acid residue-positive charge in side chain
•Length of typical Double DNA helix in mammalian calculated as 2.2
meters
•Cells-Eukaryotes & Prokaryotes
•Eukaryotes DNA localized inside the nucleus
•Prokaryotes like E. Coli-defined nucleus & DNA is not scattered
throughout in cell
•DNA-negatively charged; Protein-Positive charges
•DNA held with positive charged Protein-region termed ‘Nucleoid’
•DNA in nucleoid organized in large loops held by protein
•Eukaryotes-organization is complex; DNA forms complex with
positively charged protein-histones
•Proteins acquires charge-abundance of amino acid residues lysines&
arginines
•Amino acid residue-positive charge in side chain
Histone:
•Histones rich in amino acids-Lysines& Arginines
•Histones organized-unit of eight molecules-histone octamer
•DNA-negatively charged, wrap around positively charged histone
octamerto form a structure-nucleosome
•A typical nucleosome-200 bpof DNA helix
•Nucleosome is the repeating unit of a structure in a nucleus-Chromatin
•Chromatin-thread like stained bodies in nucleus
•Nucleosome in chromatin sees as-‘beads on string’-Electron
microscope
•Histones rich in amino acids-Lysines& Arginines
•Histones organized-unit of eight molecules-histone octamer
•DNA-negatively charged, wrap around positively charged histone
octamerto form a structure-nucleosome
•A typical nucleosome-200 bpof DNA helix
•Nucleosome is the repeating unit of a structure in a nucleus-Chromatin
•Chromatin-thread like stained bodies in nucleus
•Nucleosome in chromatin sees as-‘beads on string’-Electron
microscope
•Beads-on-string structure in chromatin packaged-Chromatin fibers-
further coiled & condensed during Metaphase-Chromosomes
•Packaging of chromatin require additional set of proteins-Non-histone
Chromosomal (NHC) proteins
•Typical nucleus-region of chromatin loosely packed-Euchromatin
(transcriptionally active); chromatin densely packed-Heterochromatin
(inactive)
further coiled & condensed during Metaphase-Chromosomes
•Packaging of chromatin require additional set of proteins-Non-histone
Chromosomal (NHC) proteins
•Typical nucleus-region of chromatin loosely packed-Euchromatin
(transcriptionally active); chromatin densely packed-Heterochromatin
(inactive)
The search for genetic material
Transforming Principle:
•Principle or factor responsible for heredity in organism
•1928, Frederick Griffith-series of experiments with Streptococcus
pneumonia –bacteria causing Pneumonia
•Observed change in the physical form of bacteria-transformation
•Streptococcus pneumonia-grown in culture plate-two different
colonies observed
i.Smooth colony-S Strain: Mucous/ polysaccharide coat-Virulent
ii.Rough colony-R Strain: Does not have mucous coat-Non virulent
•Mice infected with S strain S. pneumonia (virulent)-die, pneumonia
•Mice infected with R strain S. pneumonia (non virulent)-survived
S-Strain Injected into mice Mice Die
R-Strain Injected into mice Mice live
Transforming Principle:
•Principle or factor responsible for heredity in organism
•1928, Frederick Griffith-series of experiments with Streptococcus
pneumonia –bacteria causing Pneumonia
•Observed change in the physical form of bacteria-transformation
•Streptococcus pneumonia-grown in culture plate-two different
colonies observed
i.Smooth colony-S Strain: Mucous/ polysaccharide coat-Virulent
ii.Rough colony-R Strain: Does not have mucous coat-Non virulent
•Mice infected with S strain S. pneumonia (virulent)-die, pneumonia
•Mice infected with R strain S. pneumonia (non virulent)-survived
S-Strain Injected into mice Mice Die
R-Strain Injected into mice Mice live
•Griffith performed 2
nd
set experiment
•Killed bacteria-heating bacteria
•Heat killed S strain when injected-Mice survived
•Mixture of heat killed S and live R bacteria when injected-Mice died
S-Strain
(Heat killed)
+
R strain
(live)
Injected into mice Mice Die
S-Strain
(Heat Killed)
Injected into mice Mice live
•Conclusion of experiment-R strain bacteria transformed by heat
killed S strain
•‘Transforming principle’-transferred from heat killed S strain which
enabled R strain to synthesize a smooth polysaccharide coat-Virulent-
transfer of genetic material
•Biochemical nature of genetic material-not defined
nd
set experiment
•Killed bacteria-heating bacteria
•Heat killed S strain when injected-Mice survived
•Mixture of heat killed S and live R bacteria when injected-Mice died
S-Strain
(Heat killed)
+
R strain
(live)
Injected into mice Mice Die
S-Strain
(Heat Killed)
Injected into mice Mice live
•Conclusion of experiment-R strain bacteria transformed by heat
killed S strain
•‘Transforming principle’-transferred from heat killed S strain which
enabled R strain to synthesize a smooth polysaccharide coat-Virulent-
transfer of genetic material
•Biochemical nature of genetic material-not defined
Biochemical Characterization of Transforming Principle
•Initially genetic material thought as protein until the work of Oswald
Avery, Colin MacLeod & MaclynMcCarty-1933-44
•They work to determine-biochemical nature of ‘transforming principle’-
Griffith’s experiment
•Their work included-purification of biochemical-protein, DNA, RNA
from heat killed S strain cells-determine the factor responsible of
transformation of R strain to S strain
•Work concluded-DNA from heat killed S stain bacteria cause R
bacteria to transform
•When protein digesting enzymes (proteases) & RNA digesting enzyme
(RNases) is mixed with heat killed S bacteria-transformation of R
bacteria to S bacteria does take place
•Digestion of DNA with DNase-inhibit transformation-suggest DNA
cause transformation
•They concluded DNA as hereditary material-not biologist were
convinced
•Initially genetic material thought as protein until the work of Oswald
Avery, Colin MacLeod & MaclynMcCarty-1933-44
•They work to determine-biochemical nature of ‘transforming principle’-
Griffith’s experiment
•Their work included-purification of biochemical-protein, DNA, RNA
from heat killed S strain cells-determine the factor responsible of
transformation of R strain to S strain
•Work concluded-DNA from heat killed S stain bacteria cause R
bacteria to transform
•When protein digesting enzymes (proteases) & RNA digesting enzyme
(RNases) is mixed with heat killed S bacteria-transformation of R
bacteria to S bacteria does take place
•Digestion of DNA with DNase-inhibit transformation-suggest DNA
cause transformation
•They concluded DNA as hereditary material-not biologist were
convinced
The genetic material is dna
•Alfred Hershey & Martha Chase (1952)-proved DNA as the genetic
material
•Worked with Bacteriophages (virus)-infects bacteria
•Bacteriophage attaches to bacteria & injects its genetic material into
bacteria-viral genetic material incorporates itself to bacterial DNA-
many new viral particles are formed
•Hershey & Chase worked to discover-whether ‘protein or DNA’ of
virus has entered the cell
Experiment of Hershey & Chase
•Grew some virus on medium containing radioactive phosphorus
(
32
P) & some other in medium containing radioactive sulfur (
35
S)
•DNA contains phosphorous & protein does not; likewise Sulfur is
present in protein not in DNA
•Alfred Hershey & Martha Chase (1952)-proved DNA as the genetic
material
•Worked with Bacteriophages (virus)-infects bacteria
•Bacteriophage attaches to bacteria & injects its genetic material into
bacteria-viral genetic material incorporates itself to bacterial DNA-
many new viral particles are formed
•Hershey & Chase worked to discover-whether ‘protein or DNA’ of
virus has entered the cell
Experiment of Hershey & Chase
•Grew some virus on medium containing radioactive phosphorus
(
32
P) & some other in medium containing radioactive sulfur (
35
S)
•DNA contains phosphorous & protein does not; likewise Sulfur is
present in protein not in DNA
•Virus grown in the presence of radioactive phosphorus-radioactive
DNA not radioactive protein
•Virus-presence of radioactive sulfur-radioactive protein not radioactive
DNA
•Radioactive phages-allowed to attach E. coli bacteria to continue the
infection; viral coats were removed from bacteria-agitating the culture
in blender
•Virus particles-separated from bacteria-centrifuge
Observation
•Bacteria infected with virus with radioactive DNA were radioactive,
supernatant-non radioactive
•Bacteria infected with virus with radioactive protein were not
radioactive, supernatant-radioactive
Conclusion
•DNA is the genetic material that is passed from virus to bacteria
•Protein is not the genetic material
DNA not radioactive protein
•Virus-presence of radioactive sulfur-radioactive protein not radioactive
DNA
•Radioactive phages-allowed to attach E. coli bacteria to continue the
infection; viral coats were removed from bacteria-agitating the culture
in blender
•Virus particles-separated from bacteria-centrifuge
Observation
•Bacteria infected with virus with radioactive DNA were radioactive,
supernatant-non radioactive
•Bacteria infected with virus with radioactive protein were not
radioactive, supernatant-radioactive
Conclusion
•DNA is the genetic material that is passed from virus to bacteria
•Protein is not the genetic material
Properties of genetic material (DNA vs rna)
•Experiment of Hershey & Chase-proved DNA acts as genetic material
•Subsequent studies showed RNA as genetic material of viruses
( Eg. Tobacco Mosaic Virus, QB Bacteriophage)
•A molecule to act as genetic material have following criteria:
1.It should be able to generate its replica (Replication)
2.It should chemically & structurally be stable
3.Provide the scope for slow changes (mutation)-required for evolution
4.Should be able to express itself in the form of ‘Mendelian Characters’
Replication:
•Replication refer to duplications
•Nucleic acids (DNA & RNA)-ability to duplicate due to base pairing &
Complementarity
•Other biomolecule (protein) cannot undergo duplication
•Experiment of Hershey & Chase-proved DNA acts as genetic material
•Subsequent studies showed RNA as genetic material of viruses
( Eg. Tobacco Mosaic Virus, QB Bacteriophage)
•A molecule to act as genetic material have following criteria:
1.It should be able to generate its replica (Replication)
2.It should chemically & structurally be stable
3.Provide the scope for slow changes (mutation)-required for evolution
4.Should be able to express itself in the form of ‘Mendelian Characters’
Replication:
•Replication refer to duplications
•Nucleic acids (DNA & RNA)-ability to duplicate due to base pairing &
Complementarity
•Other biomolecule (protein) cannot undergo duplication
Stability:
•Genetic material should be stable-should not change with different
stages of life cycle, age, change in physiology
•Stability of genetic material was evident with Griffith’s ‘transforming
principle’-heat killed the bacteria but didn’t destroy the properties of
genetic material-can be explained as DNA strands (complementary)
separate when heated & pair again at appropriate conditions
•RNA-2’ OH group at every nucleotide is reactive-RNA labile & easily
degradable
•RNA is known to be catalytic-reactive
•DNA-chemically less reactive & structurally more stable than RNA
which makes DNA better genetic material
•Presence of Thymine in place of Uracil-adds stability
Mutation:
•Both DNA & RNA-mutate
•RNA-unstable mutate faster; Virus with RNA genome-shorter life
span-prone to mutation & evolve
•Genetic material should be stable-should not change with different
stages of life cycle, age, change in physiology
•Stability of genetic material was evident with Griffith’s ‘transforming
principle’-heat killed the bacteria but didn’t destroy the properties of
genetic material-can be explained as DNA strands (complementary)
separate when heated & pair again at appropriate conditions
•RNA-2’ OH group at every nucleotide is reactive-RNA labile & easily
degradable
•RNA is known to be catalytic-reactive
•DNA-chemically less reactive & structurally more stable than RNA
which makes DNA better genetic material
•Presence of Thymine in place of Uracil-adds stability
Mutation:
•Both DNA & RNA-mutate
•RNA-unstable mutate faster; Virus with RNA genome-shorter life
span-prone to mutation & evolve
Protein Synthesis ( Genetic Expression):
•RNA directly codes for synthesis of protein-express characters
easily
•DNA-dependent on RNA for protein synthesis
•Protein synthesis machinery-evolved around RNA
Conclusion
•DNA & RNA-function as genetic material
•DNA being more stable preferred as storage of genetic
information
•RNA-transmission of genetic information
•RNA directly codes for synthesis of protein-express characters
easily
•DNA-dependent on RNA for protein synthesis
•Protein synthesis machinery-evolved around RNA
Conclusion
•DNA & RNA-function as genetic material
•DNA being more stable preferred as storage of genetic
information
•RNA-transmission of genetic information
Rna world
•What is the first genetic material? DNA / RNA
•Evidences suggests essential life processes like metabolism,
translation & splicing evolved around RNA
•RNA-genetic material in organism like Virus, also Catalyst of some
biochemical reactions in living systems
•But as catalyst-unstable due to reactive 2’-OH group
•DNA has evolved from RNA with some modifications
•Modifications include-double stranded nature & complementary
strand-resist the changes by evolving the process of repair
•What is the first genetic material? DNA / RNA
•Evidences suggests essential life processes like metabolism,
translation & splicing evolved around RNA
•RNA-genetic material in organism like Virus, also Catalyst of some
biochemical reactions in living systems
•But as catalyst-unstable due to reactive 2’-OH group
•DNA has evolved from RNA with some modifications
•Modifications include-double stranded nature & complementary
strand-resist the changes by evolving the process of repair
replication
•“It is the process by which DNA produces the exact copies of the
original DNA."
•Replication takes place during S-phase of cell cycle
•Proposed by Watson and Crick following the double helical structure
•Watson & Crick’s original statement (1953) states:
“It was not escaped our notice that the specific pairing we have
postulated immediately suggests a possible copying mechanism for the
genetic material”
•They suggested DNA replication-Semi-conservative method
•Described process-two DNA strand separate & act as template for
synthesis of new complementary DNA strand-complementary base
parind
•New DNA molecule formed have-one parental DNA strand & one
newly synthesized daughter DNA strand
•“It is the process by which DNA produces the exact copies of the
original DNA."
•Replication takes place during S-phase of cell cycle
•Proposed by Watson and Crick following the double helical structure
•Watson & Crick’s original statement (1953) states:
“It was not escaped our notice that the specific pairing we have
postulated immediately suggests a possible copying mechanism for the
genetic material”
•They suggested DNA replication-Semi-conservative method
•Described process-two DNA strand separate & act as template for
synthesis of new complementary DNA strand-complementary base
parind
•New DNA molecule formed have-one parental DNA strand & one
newly synthesized daughter DNA strand
Dna Replication Pattern
Conservativereplication
wouldleaveintacttheoriginalDNAmolecule
andgenerateacompletelynewmolecule.
Dispersivereplication
wouldproducetwoDNAmoleculeswithsections
ofbotholdandnewDNAinterspersedalongeach
strand.
Semiconservativereplication
wouldproducemoleculeswithbotholdand
newDNA,buteachmoleculewouldbe
composedofoneoldstrandandonenew
strand.
wouldleaveintacttheoriginalDNAmolecule
andgenerateacompletelynewmolecule.
Dispersivereplication
wouldproducetwoDNAmoleculeswithsections
ofbotholdandnewDNAinterspersedalongeach
strand.
Semiconservativereplication
wouldproducemoleculeswithbotholdand
newDNA,buteachmoleculewouldbe
composedofoneoldstrandandonenew
strand.
Matthew Meselson and Franklin Stahl experiment
•They grew E. colion
15
NH
4Cl culture medium.
15
N is the heavy isotope
of nitrogen-gets incorporated in the new DNA synthesized
•Both strands of DNA have
15
N (
15
N
15
N)-0 Minute
•DNA extracted-subjected to CsCldensity gradient centrifugations after
every definite interval
•Samples separated independently in CsClgradients based on difference
in their densities
•Bacteria is then shifted to
14
NH
4Cl culture medium-
14
N is light isotope of
Nitrogen
•DNA extraction, centrifugation with CsCl-after 20 minutes-First
generation (I)
•DNA extracted from first generation (
15
N to
14
N) -Hybrid/ Intermediate
type of DNA (
15
N
14
N)-20 Minute
•Second generation-equal amount of light DNA (
14
N
14
N) and hybrid
DNA (
15
N
14
N) are formed.-40 Minute
•They grew E. colion
15
NH
4Cl culture medium.
15
N is the heavy isotope
of nitrogen-gets incorporated in the new DNA synthesized
•Both strands of DNA have
15
N (
15
N
15
N)-0 Minute
•DNA extracted-subjected to CsCldensity gradient centrifugations after
every definite interval
•Samples separated independently in CsClgradients based on difference
in their densities
•Bacteria is then shifted to
14
NH
4Cl culture medium-
14
N is light isotope of
Nitrogen
•DNA extraction, centrifugation with CsCl-after 20 minutes-First
generation (I)
•DNA extracted from first generation (
15
N to
14
N) -Hybrid/ Intermediate
type of DNA (
15
N
14
N)-20 Minute
•Second generation-equal amount of light DNA (
14
N
14
N) and hybrid
DNA (
15
N
14
N) are formed.-40 Minute
DNA replication-introduction
•Enzymes are involved in the process of replication
•Action of replication take place with high accuracy to avoid mutation
•DNA polymerase-main enzyme and is DNA dependent since DNA
act as template & catalyse polymerization of deoxyribonucleotides
•E. Coli–4.6 x 10
6
bp, replication completes-38 minutes
•Polymerase-have to be fast & should catalyse reaction with high
degree accuracy
•Deoxyribonucleoside triphosphate-substrate & provide energy for
polymerization
•Enzymes are involved in the process of replication
•Action of replication take place with high accuracy to avoid mutation
•DNA polymerase-main enzyme and is DNA dependent since DNA
act as template & catalyse polymerization of deoxyribonucleotides
•E. Coli–4.6 x 10
6
bp, replication completes-38 minutes
•Polymerase-have to be fast & should catalyse reaction with high
degree accuracy
•Deoxyribonucleoside triphosphate-substrate & provide energy for
polymerization
Dna replication
Components Essential For Replication:
1.Enzymes-Helicase, Topoisomerase, Primase, DNA Polymerase,
DNA ligase, Repair enzymes
2.Nucleotides-Deoxyribonucleotides
3.Energy-ATP & Deoxyribonucleoside triphosphate
•The steps involved in the process of DNA replication includes:
1.Origination of replication
2.Unwinding of DNA molecule
3.Formation of RNA primer
4.Synthesis of complementary strand
5.Proof reading and DNA repair
Components Essential For Replication:
1.Enzymes-Helicase, Topoisomerase, Primase, DNA Polymerase,
DNA ligase, Repair enzymes
2.Nucleotides-Deoxyribonucleotides
3.Energy-ATP & Deoxyribonucleoside triphosphate
•The steps involved in the process of DNA replication includes:
1.Origination of replication
2.Unwinding of DNA molecule
3.Formation of RNA primer
4.Synthesis of complementary strand
5.Proof reading and DNA repair
1.Origination of Replication
•Replication starts at a small opening of DNA helix as a complete
DNA cannot be separated
•initiation points or origin-hydrogen bond between two bases
breaks
2. Unwinding of DNA molecule
•Enzyme Helicase-unwinds & separate two strand
•Involves breaking weak hydrogen bond
•Energy required-ATP
•Enzyme Topoisomerase-cut and reseal one strand of DNA strand
helping separation of interwinedstrand of long DNA molecule
•Leads to formation of Y-shaped replication fork
3. Formation of RNA primer
•RNA primer-short segment of RNA nucleotide formed at DNA
template before replication begins
•Initiates the formation of new complementary DNA strand-3’ end
•Replication starts at a small opening of DNA helix as a complete
DNA cannot be separated
•initiation points or origin-hydrogen bond between two bases
breaks
2. Unwinding of DNA molecule
•Enzyme Helicase-unwinds & separate two strand
•Involves breaking weak hydrogen bond
•Energy required-ATP
•Enzyme Topoisomerase-cut and reseal one strand of DNA strand
helping separation of interwinedstrand of long DNA molecule
•Leads to formation of Y-shaped replication fork
3. Formation of RNA primer
•RNA primer-short segment of RNA nucleotide formed at DNA
template before replication begins
•Initiates the formation of new complementary DNA strand-3’ end
4. Synthesis of New Complementary strand:
•After formation of RNA primer-new nucleotides are added to RNA
primer-DNA polymerase-add nucleotides in 5’3’ only
•New complementary strand proceeds from 3’ -OH terminus of the primer
causing extension in 5’3’ direction
•Building blocks for polymerization-deoxyribonucleotidetriphosphate
(dATP, dTTP, dGTP, dCTP)
•Replication proceeds in 5’3’ in one strand having template polarity
3’5’-leading strand-Polymerase enzyme produce continuous strand
since replication is continuous
•Opposite parental DNA strand with template polarity 5’3’–antiparallel;
DNA polymerase produces short sections of DNA strand (5’3’) -these
short fragments of DNA strand-lagging strand/ discontinuous strand
& fragments are called Okazaki fragments
•Okazaki fragments/ DNA fragments-joined by DNA ligase
•After formation of RNA primer-new nucleotides are added to RNA
primer-DNA polymerase-add nucleotides in 5’3’ only
•New complementary strand proceeds from 3’ -OH terminus of the primer
causing extension in 5’3’ direction
•Building blocks for polymerization-deoxyribonucleotidetriphosphate
(dATP, dTTP, dGTP, dCTP)
•Replication proceeds in 5’3’ in one strand having template polarity
3’5’-leading strand-Polymerase enzyme produce continuous strand
since replication is continuous
•Opposite parental DNA strand with template polarity 5’3’–antiparallel;
DNA polymerase produces short sections of DNA strand (5’3’) -these
short fragments of DNA strand-lagging strand/ discontinuous strand
& fragments are called Okazaki fragments
•Okazaki fragments/ DNA fragments-joined by DNA ligase
5. Proof reading and DNA repair:
•Accuracy of base pairing is essential to avoid mutation
•Any error one in 10,000 is also rectified by removing wrong base &
replacing correct base
•DNA polymerase-remove & replace any wrong base in the newly
synthesized strand
•Repair enzyme (Nuclease)-cut off any defection in strand
•Proof reading ensures identical DNA strand
•Accuracy of base pairing is essential to avoid mutation
•Any error one in 10,000 is also rectified by removing wrong base &
replacing correct base
•DNA polymerase-remove & replace any wrong base in the newly
synthesized strand
•Repair enzyme (Nuclease)-cut off any defection in strand
•Proof reading ensures identical DNA strand
Dna replication-Quiz
transcription
•Process of copying genetic information from one strand of DNA into
single stranded RNA
•Depends upon the principle of complementarity(C::G & A::U), in RNA
strand in place of Thymine, Uracil is present
•Only a segment of DNA and only one of the strand of DNA is copied into
RNA, so it is very necessary to define the region on DNA that should be
transcribed
•Both DNA strands cannot be transcribed into RNA due to following
reason:
1.If both strand act as template, they produce 2 different RNA molecule
with different sequence (complementarity); they code for two different
protein since the amino acids of both RNA transcribed are different &
one segment of DNA will code for two different proteins-complicate
genetic information
2.Two RNA molecules produced simultaneously-complementary-double
stranded RNA-this prevent RNA translation to protein
•Process of copying genetic information from one strand of DNA into
single stranded RNA
•Depends upon the principle of complementarity(C::G & A::U), in RNA
strand in place of Thymine, Uracil is present
•Only a segment of DNA and only one of the strand of DNA is copied into
RNA, so it is very necessary to define the region on DNA that should be
transcribed
•Both DNA strands cannot be transcribed into RNA due to following
reason:
1.If both strand act as template, they produce 2 different RNA molecule
with different sequence (complementarity); they code for two different
protein since the amino acids of both RNA transcribed are different &
one segment of DNA will code for two different proteins-complicate
genetic information
2.Two RNA molecules produced simultaneously-complementary-double
stranded RNA-this prevent RNA translation to protein
Transcription unit
•Sequence of nucleotides in DNA that codes for a single RNA molecule,
along with the sequences necessary for its transcription
•The stretch of DNA transcribed into an RNA molecule is called
atranscription unitand encodes at least one gene
•The 3 regions in transcription unit:
i.A Promoter-initiates the process of transcription-5’ end, upstream
ii.The Structural gene-codes of RNA-RNA polymerase
iii.A Terminator-terminates transcription-3’ end, downstream
•Additional regulatory genespresent upstream & downstream to
promoter-regulate transcription
•Sequence of nucleotides in DNA that codes for a single RNA molecule,
along with the sequences necessary for its transcription
•The stretch of DNA transcribed into an RNA molecule is called
atranscription unitand encodes at least one gene
•The 3 regions in transcription unit:
i.A Promoter-initiates the process of transcription-5’ end, upstream
ii.The Structural gene-codes of RNA-RNA polymerase
iii.A Terminator-terminates transcription-3’ end, downstream
•Additional regulatory genespresent upstream & downstream to
promoter-regulate transcription
•Transcriptionis the first step ofgene expression, in which a particular
segment ofDNA is copied intoRNAby theenzyme RNA Polymerase.
•DNA base pairsof nucleotidesas a complementarylanguage that can be
converted from DNA to RNA by the action of enzymes.
•Both strands of DNA (antiparallel) is defined as structural gene in
transcription unit
•Enzyme DNA dependent RNA polymerase catalyse polymerization from
5’3’ only
•DNA strand with polarity 3’5’ act as template-template strand
•Opposite strand will have 5’3’ polarity & have base sequences exactly
same as the transcribed RNA(except thymine in place of uracil)-coding
strand
•All reference while defining transcription unit is made with coding strand
segment ofDNA is copied intoRNAby theenzyme RNA Polymerase.
•DNA base pairsof nucleotidesas a complementarylanguage that can be
converted from DNA to RNA by the action of enzymes.
•Both strands of DNA (antiparallel) is defined as structural gene in
transcription unit
•Enzyme DNA dependent RNA polymerase catalyse polymerization from
5’3’ only
•DNA strand with polarity 3’5’ act as template-template strand
•Opposite strand will have 5’3’ polarity & have base sequences exactly
same as the transcribed RNA(except thymine in place of uracil)-coding
strand
•All reference while defining transcription unit is made with coding strand
•Promoter & terminator-flank structural gene i.e., located at either
end of structural gene
•Location of promoter & terminator-polarity of Coding strand
•Promoter
i.located 5’end, upstream of the structural gene
ii.DNA sequence-provide binding site for RNA polymerase
iii.Presence defines template strand & coding strand, when promoter
position is switched with terminator, definition of coding & template
strand is reversed
•Terminator
i.Located towards 3’ end of structural gene (coding strand) ,
downstream
ii.Defines the end of transcription
end of structural gene
•Location of promoter & terminator-polarity of Coding strand
•Promoter
i.located 5’end, upstream of the structural gene
ii.DNA sequence-provide binding site for RNA polymerase
iii.Presence defines template strand & coding strand, when promoter
position is switched with terminator, definition of coding & template
strand is reversed
•Terminator
i.Located towards 3’ end of structural gene (coding strand) ,
downstream
ii.Defines the end of transcription
Transcription unit and gene
•DNA -genes located-functional unit of inheritance
•DNA sequence-codes for tRNA or rRNA& defined as gene
•DNA sequence coding polypeptide-Cistron
•Cistron-segment of DNA in the structural gene in the transcription unit
coding for polypeptide (protein) which could be monocistronic
(eukaryotes) / polycistronic (prokaryotes or bacteria)
•Monocistronicstructural gene-
1.genes which codes for only one mRNA & translates to one protein
(Eukaryotes)
2.Interrupted coding sequence i.e., genes are split
3.Coding/ expressed sequence-Exon(sequence which appear in
mature/ processed RNA)
4.Exons-interrupted by introns-do not appear in mature or processed
RNA
•Polycistronic gene-genes which synthesize one mRNA but can
translate more than one protein(Prokaryotes)
•Regulatory gene-do not code for RNA/ Protein
•DNA -genes located-functional unit of inheritance
•DNA sequence-codes for tRNA or rRNA& defined as gene
•DNA sequence coding polypeptide-Cistron
•Cistron-segment of DNA in the structural gene in the transcription unit
coding for polypeptide (protein) which could be monocistronic
(eukaryotes) / polycistronic (prokaryotes or bacteria)
•Monocistronicstructural gene-
1.genes which codes for only one mRNA & translates to one protein
(Eukaryotes)
2.Interrupted coding sequence i.e., genes are split
3.Coding/ expressed sequence-Exon(sequence which appear in
mature/ processed RNA)
4.Exons-interrupted by introns-do not appear in mature or processed
RNA
•Polycistronic gene-genes which synthesize one mRNA but can
translate more than one protein(Prokaryotes)
•Regulatory gene-do not code for RNA/ Protein
Types of rna
•Three types of RNA:
1.mRNA (messenger RNA)
2.tRNA (transfer RNA)
3.rRNA(ribosomal RNA)
•RNAs-necessary for protein synthesis
•mRNA-act as template for translation to protein
•tRNA-brings amino acids for protein synthesis & reads genetic code
•rRNA-structural & catalytic role during translation
•DNA dependent RNA polymerase-catalyze transcription
•Three types of RNA:
1.mRNA (messenger RNA)
2.tRNA (transfer RNA)
3.rRNA(ribosomal RNA)
•RNAs-necessary for protein synthesis
•mRNA-act as template for translation to protein
•tRNA-brings amino acids for protein synthesis & reads genetic code
•rRNA-structural & catalytic role during translation
•DNA dependent RNA polymerase-catalyze transcription
Components essential for transcription
•Components essential for transcription include:
1.RNA polymerase-binds to Promoter& initiates transcription
2.Nucleoside triphosphate-substrate & polymerizes in template
depended fashion (complementarity)
•During polymerization-RNA polymerase with short stretch of RNA
binds with template strand
•At terminator-nascent RNA with Polymerase falls off-Termination
The process of transcription includes:
1.Initiation
2.Elongation
3.Termination
•Components essential for transcription include:
1.RNA polymerase-binds to Promoter& initiates transcription
2.Nucleoside triphosphate-substrate & polymerizes in template
depended fashion (complementarity)
•During polymerization-RNA polymerase with short stretch of RNA
binds with template strand
•At terminator-nascent RNA with Polymerase falls off-Termination
The process of transcription includes:
1.Initiation
2.Elongation
3.Termination
Transcription in prokaryotes
•Transcription in prokaryotes include 3 steps: initiation, elongation &
termination
•RNA polymerase catalyse polymerization
•Enzyme Polymerase is capable-elongation only
•Associates with initiation factor (σsigma) for initiation process &
termination factor (ρrho) to terminate transcription
•Association of sigma & rho factor alters specificity of RNA
polymerase-initiate & terminate transcription
•mRNA transcribed does not require any processing to become active
•Prokaryotes-nucleus & cytoplasm are not separate
•Translation of mRNA begins before complete transcription of mRNA
•So, transcription & translation is coupled in prokaryotes
•Transcription in prokaryotes include 3 steps: initiation, elongation &
termination
•RNA polymerase catalyse polymerization
•Enzyme Polymerase is capable-elongation only
•Associates with initiation factor (σsigma) for initiation process &
termination factor (ρrho) to terminate transcription
•Association of sigma & rho factor alters specificity of RNA
polymerase-initiate & terminate transcription
•mRNA transcribed does not require any processing to become active
•Prokaryotes-nucleus & cytoplasm are not separate
•Translation of mRNA begins before complete transcription of mRNA
•So, transcription & translation is coupled in prokaryotes
Transcription in Bacteria
Transcription in eukaryotes
•Components essential for transcription includes:
1.RNA polymerase: RNA polymerase I, RNA polymerase II & RNA
polymerase III-nucleus of cell
2.Nucleoside triphosphate
•Structural gene of transcription unit-introns(does not appear) & exons
(appear as mRNA)
•During transcription RNA polymerase I-rRNAs(28S, 18S & 5.8S)
•RNA polymerase III-tRNA, 5S rRNA& snRNA(small nuclear RNAs)
•RNA polymerase II-precursor of mRNA-heterogeneous nuclear
RNA (hnRNA)
•Precursor RNA or nascent RNA-hnRNAwhich undergo capping &
splicing to form mRNA
•Components essential for transcription includes:
1.RNA polymerase: RNA polymerase I, RNA polymerase II & RNA
polymerase III-nucleus of cell
2.Nucleoside triphosphate
•Structural gene of transcription unit-introns(does not appear) & exons
(appear as mRNA)
•During transcription RNA polymerase I-rRNAs(28S, 18S & 5.8S)
•RNA polymerase III-tRNA, 5S rRNA& snRNA(small nuclear RNAs)
•RNA polymerase II-precursor of mRNA-heterogeneous nuclear
RNA (hnRNA)
•Precursor RNA or nascent RNA-hnRNAwhich undergo capping &
splicing to form mRNA
•Primary transcript have introns & exons-non functional-splicing
•Splicing-introns are removed & exons joined in defined order
•hnRNA-additional processing-capping & tailing
•Capping-Methyl guanosine triphosphate added to 5’ end of hnRNA
•Tailing-adenylateresidues (Poly adenine 200-300 approx) added-
3’end in template independent manner-fully processed hnRNA
•Fully processed hnRNAis called mRNA-transported out of nucleus
for translation
•Split gene arrangement & presence of introns-ancient feature of
genome
•Process of splicing-dominance of RNA world
•Splicing-introns are removed & exons joined in defined order
•hnRNA-additional processing-capping & tailing
•Capping-Methyl guanosine triphosphate added to 5’ end of hnRNA
•Tailing-adenylateresidues (Poly adenine 200-300 approx) added-
3’end in template independent manner-fully processed hnRNA
•Fully processed hnRNAis called mRNA-transported out of nucleus
for translation
•Split gene arrangement & presence of introns-ancient feature of
genome
•Process of splicing-dominance of RNA world
Genetic code
•Is the genetic information required for translation of polymer of
nucleotide to polymer of amino acids
•Information are transferred in the form of codes.
•This is ‘language’ of the genes, also known as the genetic code.
•There are 20 different amino acids that make up of proteins in the
body.
•But there are only 4 different bases in the DNA and RNA. This would
mean that one base cannot code for one amino acid since there are
20 amino acids available.
•A sequence of 3 bases seem the most probable since it can give 64
codon (4
3
) possible combination of bases, this could count for the
twenty amino acids available in organism.
•Is the genetic information required for translation of polymer of
nucleotide to polymer of amino acids
•Information are transferred in the form of codes.
•This is ‘language’ of the genes, also known as the genetic code.
•There are 20 different amino acids that make up of proteins in the
body.
•But there are only 4 different bases in the DNA and RNA. This would
mean that one base cannot code for one amino acid since there are
20 amino acids available.
•A sequence of 3 bases seem the most probable since it can give 64
codon (4
3
) possible combination of bases, this could count for the
twenty amino acids available in organism.
General features of genetic code.
•The genetic code is triplet: Each codon is made up of three specific
nucleotides. 61 codon code for amino acids & 3 codon do not code for
any amino acids & they function-Stop codon
•Genetic codon is unambiguous & Specific-one codon codes for only
one amino acids
•Genetic code is universal:each codon codes for specific amino acid
in all living organism. Eg.-UUU codes for Phenylalanine (phe)-bacteria
& human
•Genetic codons are no over lapping:Adjacent codon never shares
there nucleotides.
•Genetic code is comma less/ punctuation: There is no gap
(contiguous) between neighboring codon.
•Genetic codon is degenerative in nature:more than one codon
codes for single amino acid.
•Ex GUA, GUG, GUC,GUU codes for valine.
•AUG has dual function, codes for Methionine (met) & initiator codon
•The genetic code is triplet: Each codon is made up of three specific
nucleotides. 61 codon code for amino acids & 3 codon do not code for
any amino acids & they function-Stop codon
•Genetic codon is unambiguous & Specific-one codon codes for only
one amino acids
•Genetic code is universal:each codon codes for specific amino acid
in all living organism. Eg.-UUU codes for Phenylalanine (phe)-bacteria
& human
•Genetic codons are no over lapping:Adjacent codon never shares
there nucleotides.
•Genetic code is comma less/ punctuation: There is no gap
(contiguous) between neighboring codon.
•Genetic codon is degenerative in nature:more than one codon
codes for single amino acid.
•Ex GUA, GUG, GUC,GUU codes for valine.
•AUG has dual function, codes for Methionine (met) & initiator codon
•Genetic code:The three specific nucleotide sequence that codes
for specific amino acid present in DNA is called genetic code.
•Codon: The three specific nucleotide sequence that codes for
specific amino acid present in m-RNA is called codon.
•Anti codon: The three specific nucleotide sequence that codes for
specific amino acid present in anticodon arm of t-RNA is called
anticodon.
•Anticodons are complementary to codons.
for specific amino acid present in DNA is called genetic code.
•Codon: The three specific nucleotide sequence that codes for
specific amino acid present in m-RNA is called codon.
•Anti codon: The three specific nucleotide sequence that codes for
specific amino acid present in anticodon arm of t-RNA is called
anticodon.
•Anticodons are complementary to codons.
•Initiator codon: AUG is the initiator codon that initiates the protein
synthesis. It codes for amino acidmethionine.
•Terminator codon: Among 64 codon, three codon UAG, UGA, UAA
does not codes for any amino acids. These are called terminator codon
or non-sense codon.
•Genetic code is unambiguous:Each codon codes for specific amino
acid, and only one amino acid.
•Ex: The codon UUU codes for the amino acid phenyl alanine only.
•Genetic code is unidirectional:codon reads only in 5’ to 3’ direction.
•Wobble base:In each codon third nitrogenous base is less specific.
•Ex: GUA, GUC, GUG, GUU codes for valine.
synthesis. It codes for amino acidmethionine.
•Terminator codon: Among 64 codon, three codon UAG, UGA, UAA
does not codes for any amino acids. These are called terminator codon
or non-sense codon.
•Genetic code is unambiguous:Each codon codes for specific amino
acid, and only one amino acid.
•Ex: The codon UUU codes for the amino acid phenyl alanine only.
•Genetic code is unidirectional:codon reads only in 5’ to 3’ direction.
•Wobble base:In each codon third nitrogenous base is less specific.
•Ex: GUA, GUC, GUG, GUU codes for valine.
Mutation and genetic code
•Insertion or deletion of one or two bases changes the reading frame
from the point of insertion or deletion-Point Mutation.
•A classical example of point mutation is a change of single base pair
in the gene for beta globin chain that results in the change of amino
acid residue glutamate to valine. It results into a diseased condition
called as sickle cell anemia
•Insertion or deletion of three or its multiple bases insert or delete one
or multiple codon hence one or multiple amino acids, and reading
frame remains unaltered from that point onwards. Such mutations
are referred to as frame-shift insertion or deletion mutations.
•Insertion or deletion of one or two bases changes the reading frame
from the point of insertion or deletion-Point Mutation.
•A classical example of point mutation is a change of single base pair
in the gene for beta globin chain that results in the change of amino
acid residue glutamate to valine. It results into a diseased condition
called as sickle cell anemia
•Insertion or deletion of three or its multiple bases insert or delete one
or multiple codon hence one or multiple amino acids, and reading
frame remains unaltered from that point onwards. Such mutations
are referred to as frame-shift insertion or deletion mutations.
T RNA-the adapter molecule
•tRNA (transfer RNA)-reads code on mRNA & binds amino acids for
translation
•tRNA also called sRNA (soluble RNA)-known before genetic code
was postulated
•tRNA-specific for each amino acids
Structure:
•Single stranded RNA
•Clover leaf shaped-2D & inverted L shaped-3D
•Four double stranded regions formed by hydrogen bonding between
bases-3 loops
•Anticodon loop-three unpaired bases complementary bases (anti-
codon) to code bases of mRNA (codon)
•Amino acid accepter end-amino acids binds
•Initiator tRNA-initiation & has UAC anti codon
•No tRNA-stop codon
•tRNA (transfer RNA)-reads code on mRNA & binds amino acids for
translation
•tRNA also called sRNA (soluble RNA)-known before genetic code
was postulated
•tRNA-specific for each amino acids
Structure:
•Single stranded RNA
•Clover leaf shaped-2D & inverted L shaped-3D
•Four double stranded regions formed by hydrogen bonding between
bases-3 loops
•Anticodon loop-three unpaired bases complementary bases (anti-
codon) to code bases of mRNA (codon)
•Amino acid accepter end-amino acids binds
•Initiator tRNA-initiation & has UAC anti codon
•No tRNA-stop codon
TRNA Structure
translation
•Process of polypeptide (protein) synthesis from amino acids added to
one another as per message transcribed on mRNA
•Polymerization of amino acids to form polypeptide & order and
sequence of amino acids are defined by the sequence of bases in
mRNA
Molecules Required for Protein Synthesis:
1.RNAs (mRNA, tRNA & rRNA)
2.Ribosomes
3.Enzymes
•Process of polypeptide (protein) synthesis from amino acids added to
one another as per message transcribed on mRNA
•Polymerization of amino acids to form polypeptide & order and
sequence of amino acids are defined by the sequence of bases in
mRNA
Molecules Required for Protein Synthesis:
1.RNAs (mRNA, tRNA & rRNA)
2.Ribosomes
3.Enzymes
1.RNAs
•mRNA-template for translation, have Untranslated regions (UTR)-5’
(before start codon) & 3’ end (after stop codon)-efficient translation
•tRNA-reads code & bring amino acids related to that
•rRNA
2. Ribosomes
•Responsible for protein synthesis in cell
•Have structural RNAs & about 80 different protein
•Have two subunits large & small; inactive-separate
•Active: Small subunit attaches with mRNA to begin translation
•Large subunit-two sites (P & A) for subsequent amino acids to bind &
be close to form peptide bond
•Ribosome (23S rRNAin bacteria )-
acts as enzyme-ribozyme for
formation of peptide bond
•mRNA-template for translation, have Untranslated regions (UTR)-5’
(before start codon) & 3’ end (after stop codon)-efficient translation
•tRNA-reads code & bring amino acids related to that
•rRNA
2. Ribosomes
•Responsible for protein synthesis in cell
•Have structural RNAs & about 80 different protein
•Have two subunits large & small; inactive-separate
•Active: Small subunit attaches with mRNA to begin translation
•Large subunit-two sites (P & A) for subsequent amino acids to bind &
be close to form peptide bond
•Ribosome (23S rRNAin bacteria )-
acts as enzyme-ribozyme for
formation of peptide bond
3. Enzyme
•Enzyme involved-Amino acyl tRNA synhetase
•Used for activation of amino acid
Steps involved in the Process of Translation
1.Activation of amino acids
2.Attachment of mRNA & formation of initiation complex
3.Initiation of polypeptide chain
4.Elongation of polypeptide
5.Termination
•Enzyme involved-Amino acyl tRNA synhetase
•Used for activation of amino acid
Steps involved in the Process of Translation
1.Activation of amino acids
2.Attachment of mRNA & formation of initiation complex
3.Initiation of polypeptide chain
4.Elongation of polypeptide
5.Termination
1.Activation of Amino acids & tRNA:
•This process also called charging of tRNA/ aminoacylationof
tRNA
•Enzyme AminoacyltRNA synthaseto form aminoacyltRNA
complex
•Require Mg
+
to catalyse reaction
2. Attachment of mRNA & formation of Initiation complex:
•mRNA attach with small sub-unit of ribosome at the Start codon
•Translating mRNA-flanked with Start codon (AUG)& Stop codon
•Methionine charged tRNA having anticodon UAC pairs with AUG of
mRNA
•Large sub-unit of ribosome joins & form complex-initiation
complex
•tRNA
met
also called initiator tRNA occupies P site of large sub-unit
•This process also called charging of tRNA/ aminoacylationof
tRNA
•Enzyme AminoacyltRNA synthaseto form aminoacyltRNA
complex
•Require Mg
+
to catalyse reaction
2. Attachment of mRNA & formation of Initiation complex:
•mRNA attach with small sub-unit of ribosome at the Start codon
•Translating mRNA-flanked with Start codon (AUG)& Stop codon
•Methionine charged tRNA having anticodon UAC pairs with AUG of
mRNA
•Large sub-unit of ribosome joins & form complex-initiation
complex
•tRNA
met
also called initiator tRNA occupies P site of large sub-unit
3. Initiation of Polypeptide
•Synthesis of protein/ polypeptide
•Initiation starts with amino acid methionine carrying tRNA/ initiator RNA
when binds to P site & A site-empty with empty exposing codons of
mRNA
4. Elongation of Polypeptide
•Starts from 5’ to 3’
•2
nd
tRNA charged with appropriate amino acid with anti codon of
exposed codon of mRNA, enters the complex & binds at A site
•Peptide bond-formed between amino acid of P & A site (close)
resulting-dipeptide
•Linkage between Methionine & its carrying tRNA break & tRNA at A-site
carry di-peptide
•tRNA at A site-pulled to P-site along with mRNA-translocation
•Again A site gets exposed with another codon & relevant tRNA binds at
A site-amino acid are linked & elongation continues
•Synthesis of protein/ polypeptide
•Initiation starts with amino acid methionine carrying tRNA/ initiator RNA
when binds to P site & A site-empty with empty exposing codons of
mRNA
4. Elongation of Polypeptide
•Starts from 5’ to 3’
•2
nd
tRNA charged with appropriate amino acid with anti codon of
exposed codon of mRNA, enters the complex & binds at A site
•Peptide bond-formed between amino acid of P & A site (close)
resulting-dipeptide
•Linkage between Methionine & its carrying tRNA break & tRNA at A-site
carry di-peptide
•tRNA at A site-pulled to P-site along with mRNA-translocation
•Again A site gets exposed with another codon & relevant tRNA binds at
A site-amino acid are linked & elongation continues
5. Termination:
•Synthesis of polypeptide terminate-stop codon (UAA, UAG & UGA) of
mRNA reach A-site
•tRNA cannot recognize stop codon
•tRNA at P-site releases polypeptide in presence of releasing factors
which binds stop codon
•Dissociation of complex takes place
•Synthesis of polypeptide terminate-stop codon (UAA, UAG & UGA) of
mRNA reach A-site
•tRNA cannot recognize stop codon
•tRNA at P-site releases polypeptide in presence of releasing factors
which binds stop codon
•Dissociation of complex takes place
Regulation of gene expression
•Gene expressionis the process by which information from a gene is
used in the synthesis of afunctional gene product.
•These products are often proteins, but in non-protein coding genes
such asribosomal RNA (rRNA), transfer RNA (tRNA) or small
nuclear RNA (snRNA) genes, the product is a functional RNA
•Several steps are involved in Gene expression: Transcription, RNA
splicing, Translation & Post translation modificationof protein
•It regulates & form basis of cellular differentiation, morphogenesis,
adaptability of organism
•Regulation of gene expression in Eukaryotes is exerted at:
1.Transcriptional level (formation of primary transcript)
2.Processing level (regulation of splicing)
3.Transport of mRNA from nucleus to cytoplasm
4.Translational level
•Gene expressionis the process by which information from a gene is
used in the synthesis of afunctional gene product.
•These products are often proteins, but in non-protein coding genes
such asribosomal RNA (rRNA), transfer RNA (tRNA) or small
nuclear RNA (snRNA) genes, the product is a functional RNA
•Several steps are involved in Gene expression: Transcription, RNA
splicing, Translation & Post translation modificationof protein
•It regulates & form basis of cellular differentiation, morphogenesis,
adaptability of organism
•Regulation of gene expression in Eukaryotes is exerted at:
1.Transcriptional level (formation of primary transcript)
2.Processing level (regulation of splicing)
3.Transport of mRNA from nucleus to cytoplasm
4.Translational level
•Expression of genes can also be regulated by change in
physiological, metabolic & environmentalconditions
•Beta-galactosidase (enzyme)-present in E.Coli, catalyse hydrolysis
of Lactose (disaccharide) into galactose & glucose which is utilized
by bacteria for energy
•When lactose are not around bacteria to use as energy, bacteria will
not synthesize enzyme beta-galactosidase-explains physiological,
metabolic & environmental conditions regulate gene expression
•Development & differentiation of embryo into adult-coordinated
regulation of expression of several sets of genes
physiological, metabolic & environmentalconditions
•Beta-galactosidase (enzyme)-present in E.Coli, catalyse hydrolysis
of Lactose (disaccharide) into galactose & glucose which is utilized
by bacteria for energy
•When lactose are not around bacteria to use as energy, bacteria will
not synthesize enzyme beta-galactosidase-explains physiological,
metabolic & environmental conditions regulate gene expression
•Development & differentiation of embryo into adult-coordinated
regulation of expression of several sets of genes
•Prokaryotes-gene expression is controlled by the rate of
transcriptional initiation
•RNA polymerase attaches at promoter of transcription unit-regulated by
accessory/ regulatory protein-enable enzyme to recognize start site
(promoter)
•Regulatory protein-positively (activation) or negatively (repressor)
•Accessibility of Promoter (prokaryotic DNA) also regulated-interaction
of regulatory proteins with short DNA sequence-operators
•Operator region-adjacent to promoter region in operons(A group of
genesor a segment of DNAthatfunctionsas single transcriptionunit.)
•Most operon operator sequence binds with a repressor protein
transcriptional initiation
•RNA polymerase attaches at promoter of transcription unit-regulated by
accessory/ regulatory protein-enable enzyme to recognize start site
(promoter)
•Regulatory protein-positively (activation) or negatively (repressor)
•Accessibility of Promoter (prokaryotic DNA) also regulated-interaction
of regulatory proteins with short DNA sequence-operators
•Operator region-adjacent to promoter region in operons(A group of
genesor a segment of DNAthatfunctionsas single transcriptionunit.)
•Most operon operator sequence binds with a repressor protein
operon
•Anoperonis a functioning unit of genomic DNA containing a cluster
ofgenes/structural genesunder the control of a single promoter.The
genes aretranscribed together into an mRNA strand
andtranslatedtogether in the cytoplasm
•An operon is made up of 4 basic DNA components:
1.Promoter–a nucleotidesequence that enables a gene to be
transcribed; recognized by RNA polymerase, which initiates
transcription.
2.Regulator–Genes control the operator gene in cooperation with certain
compounds called inducers (metabolite). The regulator gene codes for
and produces a protein substance called repressor. The repressor
combines with the operator gene to repress its action.
3.Operator–a segment of DNAthat a repressor binds to;a segment
between the promoter and the genes of the operon.When repressor
protein binds to it, it obstructs the RNA polymerase from transcribing the
genes.
4.Structural genes–the genes that are co-regulated by the operon.
•Eg.-lacoperon, trpoperon, araoperon, hisoperon, valoperon
•Anoperonis a functioning unit of genomic DNA containing a cluster
ofgenes/structural genesunder the control of a single promoter.The
genes aretranscribed together into an mRNA strand
andtranslatedtogether in the cytoplasm
•An operon is made up of 4 basic DNA components:
1.Promoter–a nucleotidesequence that enables a gene to be
transcribed; recognized by RNA polymerase, which initiates
transcription.
2.Regulator–Genes control the operator gene in cooperation with certain
compounds called inducers (metabolite). The regulator gene codes for
and produces a protein substance called repressor. The repressor
combines with the operator gene to repress its action.
3.Operator–a segment of DNAthat a repressor binds to;a segment
between the promoter and the genes of the operon.When repressor
protein binds to it, it obstructs the RNA polymerase from transcribing the
genes.
4.Structural genes–the genes that are co-regulated by the operon.
•Eg.-lacoperon, trpoperon, araoperon, hisoperon, valoperon
The lac operon
•First investigated by geneticist Francois Jacob & biochemist Jacque
Monod
•Lac operon-polycistronic (prokaryotes) structural gene regulated by
common promoter & regulatory genes
•Lac operon-hydrolysis of lactose (disaccharide) to monosaccharide
galactose & glucose
•Components of lac operon:
1.Regulatory gene-igene
2.Three structural gene-z, y & a
3.Inducer-lactose (substrate)
4.Promoter-for regulatory gene (P
i) & for lac operon (P
lac)
•First investigated by geneticist Francois Jacob & biochemist Jacque
Monod
•Lac operon-polycistronic (prokaryotes) structural gene regulated by
common promoter & regulatory genes
•Lac operon-hydrolysis of lactose (disaccharide) to monosaccharide
galactose & glucose
•Components of lac operon:
1.Regulatory gene-igene
2.Three structural gene-z, y & a
3.Inducer-lactose (substrate)
4.Promoter-for regulatory gene (P
i) & for lac operon (P
lac)
•Lacoperon-found in E.Coli
•Promoter P
i-initiates the function of regulatory gene (i)
•Promoter P
lac-initiates & regulates-lac operon
•Lac operon-one regulatory gene (i-gene) & 3 structural genes (z, y & a)
•i-gene codes for repressor of lac operon which repress transcription of
structural gene
•z-gene codes for beta-galactosidase (β-gal)-hydrolysis of lactose to
galactose and glucose
•y-gene codes for permease-increases permeability of cell to β-
galactosides
•a-gene codes for transacetylase-require for metabolism of lactose
•All three structural genes are required for metabolism of lactose
•Lactose-substrate for enzyme beta-galactosidase & regulates switching
on & off of the operon-inducer
•Glucose-preferred by bacteria; in absence of glucose, lactose acts as
substrate
•Promoter P
i-initiates the function of regulatory gene (i)
•Promoter P
lac-initiates & regulates-lac operon
•Lac operon-one regulatory gene (i-gene) & 3 structural genes (z, y & a)
•i-gene codes for repressor of lac operon which repress transcription of
structural gene
•z-gene codes for beta-galactosidase (β-gal)-hydrolysis of lactose to
galactose and glucose
•y-gene codes for permease-increases permeability of cell to β-
galactosides
•a-gene codes for transacetylase-require for metabolism of lactose
•All three structural genes are required for metabolism of lactose
•Lactose-substrate for enzyme beta-galactosidase & regulates switching
on & off of the operon-inducer
•Glucose-preferred by bacteria; in absence of glucose, lactose acts as
substrate
•Inducers-lactose or allolactose
•lactose is present in medium-transported into cell through permease
•Lactose-induces activation of lacoperon
In the absence of inducer (Lactose):
•i-gene synthesize Repressor mRNAall the time which translate to
Repressor protein
•Repressor protein-binds to operator region of operon & prevents
RNA polymerase to transcribe structural genes of operon (z, y & a)
In the presence of inducer (Lactose):
•Inducer interacts with repressor and inactivates repressor-disable its
ability to bind to operator
•Enable RNA polymerase to access promoter & transcription of
structural gene proceeds to produce lac mRNA which translates to β-
galactosidase, permease & transacetylase
•Regulation of lac operon by repressor is referred as negative
regulation
•lactose is present in medium-transported into cell through permease
•Lactose-induces activation of lacoperon
In the absence of inducer (Lactose):
•i-gene synthesize Repressor mRNAall the time which translate to
Repressor protein
•Repressor protein-binds to operator region of operon & prevents
RNA polymerase to transcribe structural genes of operon (z, y & a)
In the presence of inducer (Lactose):
•Inducer interacts with repressor and inactivates repressor-disable its
ability to bind to operator
•Enable RNA polymerase to access promoter & transcription of
structural gene proceeds to produce lac mRNA which translates to β-
galactosidase, permease & transacetylase
•Regulation of lac operon by repressor is referred as negative
regulation
Human genome project
•Launched in 1990 to sequence human genome
•GENOME –The whole hereditary information of an organism that is
encoded in the DNA
•Project was based on the assumption –DNA sequence should be
different at some places since individual differ from each other
•Was called mega project due following reasons:
1.Human genome-3x 10
9
bpapprox., cost of single base pair sequence-
US $ 3, then estimated cost for project-9 billion US dollars
2.Obtained sequences if typed in book to store; with 1000 letters per
page & 1000 pages per book then, 3300 books are required to store
information of DNA sequence from a single human cell
3.Enormous data expected to generate require high speed computational
devices for data storage & retrieval and analysis
•HGP-associated with Bioinformatics
•Launched in 1990 to sequence human genome
•GENOME –The whole hereditary information of an organism that is
encoded in the DNA
•Project was based on the assumption –DNA sequence should be
different at some places since individual differ from each other
•Was called mega project due following reasons:
1.Human genome-3x 10
9
bpapprox., cost of single base pair sequence-
US $ 3, then estimated cost for project-9 billion US dollars
2.Obtained sequences if typed in book to store; with 1000 letters per
page & 1000 pages per book then, 3300 books are required to store
information of DNA sequence from a single human cell
3.Enormous data expected to generate require high speed computational
devices for data storage & retrieval and analysis
•HGP-associated with Bioinformatics
Goals of hgp
•Important goals of HGP:
i.Identify all the approximately 20,000-25,000 genes in human DNA
ii.Determine the sequences of the 3 billion chemical base pairs that
make up human DNA
iii.Store this information in databases
iv.Improve tools for data analysis
v.Transfer related technologies to other sectors, such as industries
vi.Address the ethical, legal, and social issues (ELSI) that may arise
from the project.
•HGP-13 year project which was completed by 2003; coordinated by
U.S. Department of Energy and the National Institute of Health & the
WellcomeTrust (U.K.); additional contributions came from Japan,
France, Germany, China and others
•Important goals of HGP:
i.Identify all the approximately 20,000-25,000 genes in human DNA
ii.Determine the sequences of the 3 billion chemical base pairs that
make up human DNA
iii.Store this information in databases
iv.Improve tools for data analysis
v.Transfer related technologies to other sectors, such as industries
vi.Address the ethical, legal, and social issues (ELSI) that may arise
from the project.
•HGP-13 year project which was completed by 2003; coordinated by
U.S. Department of Energy and the National Institute of Health & the
WellcomeTrust (U.K.); additional contributions came from Japan,
France, Germany, China and others
Significance of hgp
•Gave knowledge about effects of DNA variations among individuals
•Study of DNA variations in individuals can be used to diagnose,
treat and prevent many human disorders
•Learning of DNA sequence of non human organism-understanding
natural capabilities which can be applied to solve challenges in
health care, agriculture, energy production & environmental
remediation
•DNA of many non-human model were sequences-bacteria, yeast,
Caenorhabditiselegans(a free living non-pathogenic nematode),
Dorsophila(the fruit fly), plants (rice & Arabidopsis)
•Gave knowledge about effects of DNA variations among individuals
•Study of DNA variations in individuals can be used to diagnose,
treat and prevent many human disorders
•Learning of DNA sequence of non human organism-understanding
natural capabilities which can be applied to solve challenges in
health care, agriculture, energy production & environmental
remediation
•DNA of many non-human model were sequences-bacteria, yeast,
Caenorhabditiselegans(a free living non-pathogenic nematode),
Dorsophila(the fruit fly), plants (rice & Arabidopsis)
methodologies
•Two way to sequence DNA:
1.Identifying all genes that are expressed-Expressed Sequence Tags
(ESTs)
2.Sequencing whole set of genome-coding & non-coding sequence
later different regions were assigned in sequence obtained based on
its function-Sequence Annotation
•Common method employed to sequence DNA-identifying expressed
genes which require DNA fragmentation
•DNA-long polymer of nucleotide; sequencing complete DNA has
technical limitations
•Sequencing DNA require-host & vector
•Host-bacteria & yeast-cloning DNA sequence which help in
amplification of each DNA fragment
•Vector-BAC (bacterial artificial chromosome) & YAC(yeast artificial
chromosome)-introducing DNA fragment into host for cloning
•Two way to sequence DNA:
1.Identifying all genes that are expressed-Expressed Sequence Tags
(ESTs)
2.Sequencing whole set of genome-coding & non-coding sequence
later different regions were assigned in sequence obtained based on
its function-Sequence Annotation
•Common method employed to sequence DNA-identifying expressed
genes which require DNA fragmentation
•DNA-long polymer of nucleotide; sequencing complete DNA has
technical limitations
•Sequencing DNA require-host & vector
•Host-bacteria & yeast-cloning DNA sequence which help in
amplification of each DNA fragment
•Vector-BAC (bacterial artificial chromosome) & YAC(yeast artificial
chromosome)-introducing DNA fragment into host for cloning
•Steps involved includes:
1.Isolation of total DNA from a cell
2.Fragmentation of total DNA into small sizes
3.Cloning fragments of DNA in a hostwith the help of vector-
significance of cloning is to amplify each piece of DNA to sequence
DNA conveniently & with ease
4.Commonly used host-bacteria & yeast, Vector-BAC & YAC
5.Sequencing DNA fragments using automated DNA sequencer that
works on a principle developed by Frederick Sanger
6.Arrangement of DNA sequences based on overlapping regions
which is generated for sequencing
7.Alignments of DNA sequence through computer based programs
8.Annotation & assigning sequence to each chromosome
9.Assigning genetic & physical map on genome-using information on
polymorphism of restriction endonuclease recognition sites &
microsatellites (repetitive DNA sequence)
1.Isolation of total DNA from a cell
2.Fragmentation of total DNA into small sizes
3.Cloning fragments of DNA in a hostwith the help of vector-
significance of cloning is to amplify each piece of DNA to sequence
DNA conveniently & with ease
4.Commonly used host-bacteria & yeast, Vector-BAC & YAC
5.Sequencing DNA fragments using automated DNA sequencer that
works on a principle developed by Frederick Sanger
6.Arrangement of DNA sequences based on overlapping regions
which is generated for sequencing
7.Alignments of DNA sequence through computer based programs
8.Annotation & assigning sequence to each chromosome
9.Assigning genetic & physical map on genome-using information on
polymorphism of restriction endonuclease recognition sites &
microsatellites (repetitive DNA sequence)
Salient features of human genome
•The human genome contains 3164.7 million nucleotide bases.
•The average gene consists of 3000 bases, but sizes vary greatly,
with the largest known human gene being dystrophinat 2.4 million
bases.
•The total number of genes is estimated at 30,000–much lower than
previous estimates of 80,000 to 1,40,000 genes. Almost all (99.9 per
cent) nucleotide bases are exactly the same in all people.
•The functions are unknown for over 50 per cent of discovered genes.
•Less than 2 per cent of the genome codes for proteins.
•Repeated sequences make up very large portion of the human
genome.
•The human genome contains 3164.7 million nucleotide bases.
•The average gene consists of 3000 bases, but sizes vary greatly,
with the largest known human gene being dystrophinat 2.4 million
bases.
•The total number of genes is estimated at 30,000–much lower than
previous estimates of 80,000 to 1,40,000 genes. Almost all (99.9 per
cent) nucleotide bases are exactly the same in all people.
•The functions are unknown for over 50 per cent of discovered genes.
•Less than 2 per cent of the genome codes for proteins.
•Repeated sequences make up very large portion of the human
genome.
•Repetitive sequences are stretches of DNA sequences that are
repeated many times, sometimes hundred to thousand times. They
are thought to have no direct coding functions, but they shed light on
chromosome structure, dynamics and evolution.
•Chromosome 1 has most genes (2968), and the Y has the fewest
(231).
•Scientists have identified about 1.4 million locations where single
base DNA differences (SNPs –single nucleotide polymorphism,
pronounced as ‘snips’) occur in humans. This information promises
to revolutionize the processes of finding chromosomal locations for
disease-associated sequences and tracing human history.
repeated many times, sometimes hundred to thousand times. They
are thought to have no direct coding functions, but they shed light on
chromosome structure, dynamics and evolution.
•Chromosome 1 has most genes (2968), and the Y has the fewest
(231).
•Scientists have identified about 1.4 million locations where single
base DNA differences (SNPs –single nucleotide polymorphism,
pronounced as ‘snips’) occur in humans. This information promises
to revolutionize the processes of finding chromosomal locations for
disease-associated sequences and tracing human history.
Applications and future challenges
•Knowledge of human genome sequence and researches will lead us to
understand biological systems in coming decades.
•HG sequence will enable a complete new approach to biological
research
•With whole genome sequences & new high-throughput technologies, we
can approach questions systematically and on a much broader scale
•It will be possible to understand how enormous number of genes and
proteins work together in interconnected networks in the chemistry of life.
•HG sequencing is an enormous task which will require the expertise and
creativity of tens of thousands of scientists from varied disciplines in both
the public and private sectors worldwide.
•Knowledge of human genome sequence and researches will lead us to
understand biological systems in coming decades.
•HG sequence will enable a complete new approach to biological
research
•With whole genome sequences & new high-throughput technologies, we
can approach questions systematically and on a much broader scale
•It will be possible to understand how enormous number of genes and
proteins work together in interconnected networks in the chemistry of life.
•HG sequencing is an enormous task which will require the expertise and
creativity of tens of thousands of scientists from varied disciplines in both
the public and private sectors worldwide.
Dna fingerprinting
•DNA fingerprinting is a technique to find out variations in individuals
of a population at DNA level & was first developed by Dr. Alec Jeffrey
•Works on the principle of polymorphismin DNA sequence (A DNA
polymorphism is any difference in the nucleotide sequence between
individuals. These differences can be single base pair changes,
deletions, insertions, or even changes in the number of copies of a
given DNA sequence)
•DNA finger printing involves identifying differences in a specific short
nucleotide repeats called Variable Number Tandem Repeats
(VNTR), that vary in number from person to person & are inherited
•The VNTRs of two person may be of same length and sequence at
certain sites but vary at others
•DNA fingerprinting is a technique to find out variations in individuals
of a population at DNA level & was first developed by Dr. Alec Jeffrey
•Works on the principle of polymorphismin DNA sequence (A DNA
polymorphism is any difference in the nucleotide sequence between
individuals. These differences can be single base pair changes,
deletions, insertions, or even changes in the number of copies of a
given DNA sequence)
•DNA finger printing involves identifying differences in a specific short
nucleotide repeats called Variable Number Tandem Repeats
(VNTR), that vary in number from person to person & are inherited
•The VNTRs of two person may be of same length and sequence at
certain sites but vary at others
1.Extraction & Isolation-DNA is extracted & isolated from the cells in
a high speed, refrigerated centrifuge
2.Amplification-Many copies of the extracted DNA are made by
polymerase chain reaction (PCR)
3.Restriction digestion-DNA is cut into fragments with restriction
endonuclease enzyme into precise reproducible sequence
4.Separation of DNA sequences-The cut DNA fragments are
introduced & passed through electrophoresis set up containing
agarose polymer gel; the separated fragments can be visualized by
staining them with dye that shows fluorescence under ultraviolet
radiation
5.Southern Blotting-The separated DNA sequences are transferred
into nitrocellulose or nylon membrane by placing over the gel
6.Hybridization-The nylon membrane is immersed in a bath of
radioactive probes (DNA segment of known sequence); these probes
target a specific nucleotide sequence that is complementary to them
7.Autoradiography-The nylon membrane is pressed on to a x-ray
film and dark bands develop at the probe sites which resemble the
bar codes
a high speed, refrigerated centrifuge
2.Amplification-Many copies of the extracted DNA are made by
polymerase chain reaction (PCR)
3.Restriction digestion-DNA is cut into fragments with restriction
endonuclease enzyme into precise reproducible sequence
4.Separation of DNA sequences-The cut DNA fragments are
introduced & passed through electrophoresis set up containing
agarose polymer gel; the separated fragments can be visualized by
staining them with dye that shows fluorescence under ultraviolet
radiation
5.Southern Blotting-The separated DNA sequences are transferred
into nitrocellulose or nylon membrane by placing over the gel
6.Hybridization-The nylon membrane is immersed in a bath of
radioactive probes (DNA segment of known sequence); these probes
target a specific nucleotide sequence that is complementary to them
7.Autoradiography-The nylon membrane is pressed on to a x-ray
film and dark bands develop at the probe sites which resemble the
bar codes
Steps involved in DNA finger printing technology.
Let us understand dna fingerprinting !!!!
•http://www.pbslearningmedia.org/resource/tdc02.sci.life.gen.createdn
afingerprint/create-a-dna-fingerprint/
•http://www.pbslearningmedia.org/resource/tdc02.sci.life.gen.createdn
afingerprint/create-a-dna-fingerprint/
Application of DNA finger printing technology.
1.It is used to identify criminals and rapist.
2.To solve parental dispute.
3.To solve immigrant problems.
4.To identify dead bodies of soldiers died in wars.
5.To identify dead bodies of person died at accidents and bomb
blast.
6.To identify racial groups to rewrite biological evolution
7.To detect inheritable disorders.
8.To detect donor cell in case of transplantation.
1.It is used to identify criminals and rapist.
2.To solve parental dispute.
3.To solve immigrant problems.
4.To identify dead bodies of soldiers died in wars.
5.To identify dead bodies of person died at accidents and bomb
blast.
6.To identify racial groups to rewrite biological evolution
7.To detect inheritable disorders.
8.To detect donor cell in case of transplantation.
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